Chemical constituents from Syneilesis aconitifolia (II) / 中草药

Objective: To study the chemical constituents from Syneilesis aconitifolia. Methods: The chemical constituents were isolated by silica gel column chromatography and HPLC, and its structure were identified by their spectral data and physicochemical properties analysis. Results: Eighteen compounds were isolated from methanol extract ethyl acetate extracts of S.aconitifolia with the structures identified as 3β-angeloyoxy-eremophil-6-en-8-oxo-12,15β-diacid (1), quercetin-3-O-α-L-rhamnoside (2), triacontanol (3), 8β-methoxyeremophil-3,7(11)-diene-8α,12(6α,15)-dilactone (4), 3,4-dihydroxybenzoic acid (5), 8-oxo-eremophil-6,9-dien-12-oic acid (6), 8βH-eremophil-3,7(11)-dien-12,8α(14,6α)-diolide (7), 8αH-6α,10β-dihydroxyeremophilenolide (8), pinoresinol (9), 6β,8β,10β-trihydroxyeremophil-7(11)-en-12,8-olide (10), 10α,15-dihydroxy-oplopan-4-one (11), 6α,15α-epoxy-1β,4β-dihydroxyeudesmane (12), caryolane-1,9β-diol (13), (-)-clovane-2,9-diol (14), cis-3-hexenyl-β-D-glucopyranoside (15), kaempferol-3-O-α-L-rhamnoside (16), quercetin-3-O-β-D-glucopyranoside(17) and (-)-oplopan-4-one-10-α-O-β-D-glucoside (18). Conclusion: Compound 1 is a new compound, named as syneilesis acid. Compounds 4-16 and 18are isolated from this plant for the first time.

Differential serum proteomic analysis of rheumatoid arthritis patients of cold-dampness arthralgia spasm syndrome / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To preliminarily study the essence of rheumatoid arthritis (RA) patients of cold-dampness arthralgia spasm syndrome (CDASS) at the protein expression level.</p><p><b>METHODS</b>Totally 24 RA patients were recruited from Department of Rheumatology, Affiliated Hospital of Nanjing University of Chinese Medicine from July 2009 to September 2010. They were assigned to the CDASS group and the dampness-heat arthralgia spasm syndrome (DHASS) group according to Chinese medicine syndrome typing, 12 in each group. The normal control group consisted of 12 healthy volunteers from the Health Examination Center, Affiliated Hospital of Nanjing University of Chinese Medicine. The serum proteins were compared between the CDASS group and the normal control group/the DHASS group respectively using two-dimensional gel electrophoresis. The common differential protein spots of CDASS were analyzed by mass spectrometry. The SwissProt database was inquired using Mascot Software to identify differential proteins.</p><p><b>RESULTS</b>There were 81 differential protein spots between the CDASS group and the normal control group. There were 45 differential protein spots between the CDASS group and the DHASS group. Thirteen protein spots were found to be higher or lower in protein expression quantity of the CDASS group when compared with those of the other two groups. Nine differential protein spots were identified by mass spectrometry and database retrieval. It's suggested that these proteins were most likely to be related with inhibition of cellular events, such as cell proliferation, cell differentiation, and so on.</p><p><b>CONCLUSION</b>4.1 protein and DLC-1 protein were of potential significance in the diagnosis, prognostic markers, or treatment targets of RA patients of CDASS, which also provided evidence for further studies on the essence of CDASS.</p>

Proteomic study of gallbladder cancer, with special reference on the expression and significance of annexin A3 / 中华病理学杂志

<p><b>OBJECTIVE</b>To explore the potential molecular targets for diagnosis and treatment of gallbladder cancer by analyzing and comparing the proteomes expressed in human gallbladder cancer and benign gallbladder tissues.</p><p><b>METHODS</b>The proteins expressed were analyzed using two-dimensional gel electrophoresis. The differentially expressed proteins in tumors were also analyzed by mass spectrometry (MS). AnnexinA3 expression was examined by streptavidin peroxidase immunohistochemical technique on paraffin-embedded tissue sections from 50 patients of gallbladder cancer and 38 cases of chronic eholecystitis.</p><p><b>RESULTS</b>Protein extracts of individual sample in each type of tissues were separated on two-dimensional gels. There were forty six differentially expressed proteins in the tissue samples of gallbladder cancer. Seventeen proteins were successfully identified by MS, in which nine proteins were overexpressed in tumors and the other eight proteins were underexpressed. The positive expression rates of annexinA3 in gallbladder cancer was significantly higher than that in chronic cholecystitis, and the difference was statistically significant (74.0% vs 21.1%, P < 0.01). In the gallbladder cancer, no correlation was obtained between annexinA3 and age, gender or histologicl type (P > 0.05), but overexpression of annexinA3 correlated significantly with those cases with a lower histological grading (40.0% vs 82.5%, P < 0.05); lymph node or distant metastasis (40.9% vs 100%, P < 0.05); or a shorter survival time after operation (50.0% vs 93.8%, P < 0.05).</p><p><b>CONCLUSIONS</b>Significant discrepancies in protein expression exist among gallbladder cancer and benign gallbladder tissues. AnnexinA3 plays an important role in the initiation and progression of human gallbladder cancer.</p>

Comparative proteomic analysis of human gallbladder carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To find out potential molecular targets for gallbladder carcinoma diagnosis and treatment by analyzing and comparing the proteins expressed in human gallbladder carcinoma tissue and benign gallbladder tissue.</p><p><b>METHODS</b>Proteomic analysis of 6 human gallbladder carcinoma tissues and 6 benign gallbladder tissues was carried out. Total proteins of the carcinoma tissue and benign gallbladder tissue were separated by two-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were analyzed and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Immunohistochemistry was used to examine the expression of PEBP1 protein in an independent series of samples.</p><p><b>RESULTS</b>Protein extracts of individual samples in each type of tissues were separated on two-dimensional gels. There were forty six differentially expressed proteins in the gallbladder carcinom tissues. Seventeen proteins were successfully identified by MS, in which nine proteins were overexpressed in tumors while the other eight proteins were underexpressed. The increased level of PEBP1 protein in gallbladder carcinoma was further confirmed by immunohistochemical analysis.</p><p><b>CONCLUSION</b>Seventeen differentially expressed proteins were successfully characterized by comparative proteomic analysis. Those results may provide scientific foundation for screening the molecular biomarkers which can be used in diagnosis and treatment of gallbladder carcinoma, as well as to improve its prognosis and provide a new clue for carcinogenesis research of gallbladder carcinoma.</p>

Establishment of a cell-based high-throughput screening model for PPARdelta agonists / 生物工程学报

To establish a new high-throughput screening model for the agonist of PPARdelta, PPARdelta gene was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR), and subcloned to pGEM-T Vector for sequencing, then the PPARdelta fragment was excised by restriction enzymes, and inserted into pTARGET Vector to construct expression vector pTARGET-ppARdelta. Insert three copies of PPRE into pGl3-promoter vector to construct expression vector pGl3-PPRE x 3-luc. The vector pTARGET-ppARdelta was transiently cotransfected with pGl3-PPRE x 3-luc into different cell lines to assay the expression levels of luciferase. The PPARdelta agonist screening model was established and optimized. Bezafibrate and linoleic acid can induce the expression of luciferase significantly and in a dose-dependent manner. This method can be used for high throughput screening for the agonist of PPARdelta, which might become lead compounds for new anti-atheroscleriosis or anti-adiposity drugs.

Cloning and characterization of a novel glutathione transferase gene from Penicillium chrysogenum / 生物工程学报

Glutathione transferases (GSTs) are a family of multifunctional proteins that mainly catalyze the conjugation of intracellular glutathione (GSH) to a wide variety of endogenous and exogenous electrophilic compounds. GSTs play important roles in stress tolerance and in the detoxification metabolism in organisms. A novel GST gene, Pc gstB, was cloned from penicillin producing fungus Penicillium chrysogenum using RT-PCR. The open reading frame (ORF) of Pc gstB was 651 bp and encoded a peptide of 216 residues. The deduced amino acids sequence had conserved GST domain and showed 65% identity to the characterized Aspergillus fumigutus gstB. The entire ORF of Pc gstB was inserted into vector pTrc99A and transformed into Escherichia coli DH5alpha. Recombinant PcGstB was overexpressed and its GST activity toward substrate 1-chloro-2,4-dinitrobenzene (CDNB) was validated.